Life Science

Antibody Explorer
Cell Culture
Cell Culture Capabilities
Helpful Resources
New Products
Product Lines
Antibody Expression
Clasic Media & Salts
Cytogenetics
Embryo Culture
Gene Therapy
Primary Human Culture
Protein Expression/CHO
Reagents & Supplements
Sera
Serum Free Media
Stem Cells
Vaccine Production
Cell Signaling
Custom Synthesis
Drug Discovery
Molecular Biology
Plant Biotechnology
Proteomics
Life Science Quarterly

 "; } else { print "
"; } ?> Stemline™ Media and Cytokines
  

Stemline™ Hematopoietic Stem Cell Expansion Medium
Stemline™ Methylcellulose Medium
Hematopoietic Cytokines

Stemline™ Hematopoietic Stem Cell Expansion Medium

The use of human hematopoietic stem cells (HSC) isolated from umbilical cord blood as a source of cellular reconstitution following high-dose chemotherapy is now a common therapeutic modality for the treatment of malignancy. However, due to the low yield of HSC from cord blood, HSC transfusion is most effective in children and has limited application in adults. In order to obtain sufficient numbers of cells for applications of this therapeutic approach in adults, ex vivo expansion has been utilized to ensure successful engraftment and minimize the short-term effects of neutropenia and thrombocytopenia. To this end, we have developed a serum-free, animal protein-free medium, Stemline™ Hematopoietic Stem Cell Expansion Medium for the optimal expansion of cord blood HSC.

Stemline™ Hematopoietic Stem Cell Expansion Medium has been developed to promote growth of
CD34+ hematopoietic stem cells derived from bone marrow, cord blood or mobilized peripheral blood. This medium supports high viable cell densities. The elimination of serum reduces performance variability in the medium and eliminates safety risks associated with possible adventitious agents in serum.

back to top

Prod. # Product Name Description Application
S 0189 Stemline™ Hematopoietic Stem Cell Expansion Medium Liquid
Sterile filtered
The medium does not contain antibiotics or cytokines. Medium must be supplemented with desired cytokines and/or antibiotics. Human serum albumin is the only human origin material. No other animal-derived components. Cell culture tested with normal human bone marrow CD34+ cells in a 14-day culture expansion. Medium was supplemented with 100 ng/ml each of SCF (Stem Cell Factor, S7901), G-CSF (Granulocyte-Colony Stimulating Factor, G0407) and TPO (Thrombopoietin, T1568) for cell expansions. 		Stemline™ HSC Expansion Medium shows excellent expansion of total nucleated cells (TNC), committed progenitors (GM-CFC) and primitive, high proliferative potential progenitors (HPP-CFC).

For clinical-scale studies, cord blood CD34+ cells were cultured for 7 days in 100 ml Teflon® culture bags containing 50 ml of culture medium, plus cytokines. Cells were harvested from these bags and a 5 ml aliquot was transferred to a second 100 ml Teflon® bag containing 50 ml of selected medium, plus cytokines, and cultured for an additional 7-day period. At the end of both culture protocols, cells were harvested and assayed for their ability to promote the growth of cord blood CD34+ cells. The data are summarized in this Data Sheet.
back to top

Stemline™ Methylcellulose Medium

Methylcellulose-supplemented Iscove's Modified Dulbecco's Medium (IMDM) forms a semi-solid growth medium that, when supplemented with cytokines, supports the growth and differentiation of hematopoietic progenitor cells and allows the clonal progeny of a single cell to form a distinct cluster or colony.  Methylcellulose based, semi-solid culture media are routinely used for the enumeration and evaluation of stem cell-derived progeny which typically manifest as Colony Forming Units (CFU) or Burst Forming Units (BFU).

The Stemline Methylcellulose Media family of products are manufactured under tightly controlled conditions using rigorously-qualified raw materials to ensure reliable and predictable results from the culture of human and murine hematopoietic cells.  This group of products will support the growth of granulocyte colonies (CFU-G), macrophage colonies (CFU-M), granulocyte/macrophage colonies (CFU-GM), erythroid colonies (BFU-E and CFU-E), and pluripotent mixed colonies (CFU-GEMM).

back to top

Prod. # Product Name Description Application
S 3562 Stemline™ Complete Methylcellulose Medium with Erythropoietin, Human
  • Liquid, frozen
  • Aseptically processed
  • Cell culture tested in a 12 to 16 day assay, using normal human CD 34 positive progenitor cells.
    		•  Use for culture of human hematopoietic cells.

    Supports the following colony types: CFU-G; CFU-M; CFU-GM; BFU-E; CFU-E and CFU-GEMM.

    Recommended for LTC-IC assays of purified populations of very primitive human clonogenic, hematopoietic cells.
    S 3687 Stemline™ Complete Methylcellulose Medium without Erythropoietin, Human
  • Liquid, frozen
  • Aseptically processed
  • Cell culture tested in a 12 to 16 day assay, using normal human CD 34 positive progenitor cells.
    •      		Same as S 3562, except that erythropoietin has not been added.

    Use for culture of human hematopoietic cells.

    Supports the following colony types: CFU-G; CFU-M; and CFU-GM.

    Recommended for LTC-IC assays of purified populations of very primitive human clonogenic, hematopoietic cells.
    S 3812 Stemline™ Complete Methylcellulose Medium with Erythropoietin, Mouse
  • Liquid, frozen
  • Aseptically processed
  • Cell culture tested in a 7 to 10 day CFU assay, using normal mouse bone marrow cells
    		•  Use for culture of mouse  hematopoietic cells.

    Supports the following colony types: CFU-G; CFU-M; CFU-GM; BFU-E; CFU-E and CFU-GEMM.

    Recommended for assay of murine clonogenic hematopoietic progenitor cells isolated from murine bone marrow and other murine hematopoietic tissues.
    S 3937 Stemline™ Methylcellulose Medium, without Growth Factors, Human
  • Liquid, frozen
  • Without erythropoietin
  • Aseptically processed
  • Cell culture tested in a 12 to 16 day CFU assay using normal human CD 34 positive progenitor cells
    		•  Use for culture of human hematopoietic cells.

    Supports the following colony types: CFU-G; CFU-M; CFU-GM; BFU-E; CFU-E and CFU-GEMM, only if supplemented with appropriate growth factors.

    Recommended for assay of  human clonogenic, hematopoietic  progenitor cells from human blood and bone marrow.  It allows the researcher to add growth factors or cytokines of their choice.
    S 4062 Stemline™ Methylcellulose Medium, without Growth Factors, Mouse
  • Liquid, frozen
  • Without erythropoietin
  • Aseptically processed
  • Cell culture tested in a 7 to 10 day CFU assay, using normal mouse bone marrow cells.
    		•  Use for culture of mouse hematopoietic cells.

    Supports the following colony types: CFU-G; CFU-M; CFU-GM; BFU-E; CFU-E and CFU-GEMM, only if supplemented with appropriate growth factors.

    Recommended for assay of murine clonogenic hematopoietic progenitor cells isolated from murine bone marrow and other hematopoietic tissues.  It allows the researcher to add growth factors of their choice.
    back to top

    Hematopoietic Cytokines

    The following table lists a group of cytokines whose predominant actions involve the stimulation or regulation of hematopoietic cells.

    The term "colony stimulating factor" (CSF) was originally given to agents recognized to stimulate the growth of colonies containing differentiated myeloid cells from a single bone marrow-derived precursor cell plated in semi-solid agar.  Glycoproteins considered to be CSFs  include: granulocyte colony-stimulating factor (G-CSF); granulocyte-macrophage colony-stimulating factor (GM-CSF); macrophage colony-stimulating factor (M-CSF); interleukin-3 (multi-CSF or IL-3); interleukin-5 (IL-5); erythropoietin (EPO), and thrombopoietin (TPO).

    The formation and maturation of hematopoietic cells are also affected by cytokines of the structural class know as the "4-alpha-helical bundle" family.  These include stem cell factor (SCF) and  flt-3/flk-2 Ligand (FL).

    back to top
    Prod. # Description Source EC50 Value Quality Assurance Stimulated Cells
    E5627 Erythropoietin (EPO) Human;
    recombinant host:
    CHO cells    		 0.05
    to
    0.1
    units/mL    		 10 units
    Endotoxin tested
    Proliferative activity tested using the human cell line TF-1.

    		 Erythroid progenitor cells.  EPO receptors are lost during cell differentiation.
    F3422 FLT-3/FLK-2 Ligand Human;
    recombinant host:
    NSO mouse cells    		 0.5
    to
    1.0
    ng/mL    		 5  µg
    > 97% pure by (SDS-PAGE)
    Endotoxin tested
    Biological activity measured by its ability to stimulate the proliferation of an Flt3 transfected pro-B cell line.

    		 Primitive hematopoietic (multi-lineage) progenitor cells that express the flt3 receptor.
    Myeloid and pro-B cells of mouse, rat, cat, rabbit, primate and human origin.
    G0407 Granulocyte Colony-Stimulating Factor (G-CSF) Human;
    recombinant host:
    Escherichia coli    		 0.01
    to
    0.1
    ng/mL    		 2  µg
    > 95% pure by (SDS-PAGE) and N-terminal analysis.
    18.8 kDa monomer; 175 amino acids; N-termianl methionine.
    Endotoxin tested
    Proliferative activity tested using the cytokine-dependent murine NFS-60 cell line.

    		 Proliferation, differentiation, survival and activation factor for hematopoietic restricted granulocyte lineage cells.
    Neutrophils, myeloid leukemic cells, neutrophilic granulocytes from bone marrow and placental trophoblasts.
    G8160 Granulocyte Colony-Stimulating Factor (G-CSF) Mouse;
    recombinant host:
    Escherichia coli    		 0.005
    to
    0.05
    ng/mL    		 5  µg
    > 97% pure by (SDS-PAGE)
    19 kDa monomer; 179 amino acids; N-terminal methionine.
    Endotoxin tested
    Proliferative activity tested using the cytokine-dependent murine NFS-60 cell line.

    		 Proliferation, differentiation, survival and activation factor for hematopoietic restricted granulocyte lineage cells.
    Neutrophils, myeloid leukemic cells, neutrophilic granulocytes from bone marrow and placental trophoblasts.
    G5035 Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF) Human;
    recombinant host:
    Escherichia coli    		 0.02
    to
    0.2
    ng/mL    		 5  µg
    > 97% pure by (SDS-PAGE)
    14 kDa monomer; 127 amino acids; N-terminal methionine.
    Endotoxin tested
    Proliferative activity tested using human TF-1 cells

    		 Growth, differentiation, and essential survival factor for granulocyte, macrophage and eosinophil lineage cells from progenitor stage to maturity.
    Receptors on most types of myeloid progenitor cells, mature monocytes, neutrophils, eosinophils, basiophils, and dendritic cells.
    Stimulates some epithelial cells and osteoclasts.
    G0282 Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF) Mouse;
    recombinant host:
    Escherichia coli    		 0.05
    to
    0.5
    ng/mL    		 5  µg
    > 95% pure by (SDS-PAGE) and N-terminal analysis.
    14.8 kDa monomer; 125 amino acids; N-terminal methionine.
    Endotoxin tested
    Proliferative activity tested using murine DA-3 cells

    		 Growth, differentiation, and essential survival factor for granulocyte, macrophage and eosinophil lineage cells from progenitor stage to maturity.
    Receptors on most types of myeloid progenitor cells, mature monocytes, neutrophils, eosinophils, basiophils, and dendritic cells.
    Stimulates some epithelial cells and osteoclasts.
    I1646 Interleukin-3 (IL-3) Human;
    recombinant host:
    Escherichia coli    		 0.04
    to
    0.4
    ng/mL    		 10  µg
    > 97% pure by (SDS-PAGE)
    Endotoxin tested
    Proliferative activity tested using human TF-1 cells

    		 Multifunctional cytokine.
    Supports early development of multi-potential hematopoietic progenitor cells: macrophage, neutrophils, mast cells and megakaryocytes from bone marrow.
    Stimulate T-cells and induce IgG secretion from activated B-cells.
    I4144 Interleukin-3 (IL-3) Mouse;
    recombinant host:
    Escherichia coli    		 0.025
    to
    0.10
    ng/mL    		 10  µg
    > 97% pure by (SDS-PAGE)
    Endotoxin tested
    Proliferative activity tested using murine NFS-60 cells

    		 Multifunctional cytokine.
    Supports early development of multi-potential hematopoietic progenitor cells: macrophage, neutrophils, mast cells and megakaryocytes from bone marrow.
    Stimulate T-cells and induce IgG secretion from activated B-cells.
    I1395 Interleukin-6 (IL-6) Human;
    recombinant host:
    Escherichia coli    		 0.4
    to
    2.0
    ng/mL    		 10  µg
    > 97% pure by (SDS-PAGE)
    Endotoxin tested
    Proliferative activity tested using the mouse plasmacytoma cell line T1165.85.2.1.

    		 Multifunctional cytokine which acts on a wide range of cell types.
    In combination with other cytokines, it acts upon; fibroblasts, myeloid progenitor cells, T-cells, B-cells, and hepatocytes.
    I9646 Interleukin-6 (IL-6) Mouse;
    recombinant host:
    Escherichia coli    		 0.03
    to
    0.30
    ng/mL    		 5  µg
    > 97% pure by (SDS-PAGE)
    Endotoxin tested
    Proliferative activity tested using the mouse plasmacytoma cell line T1165.85.2.1.

    		 Multifunctional cytokine which acts on a wide range of cell types.
    In combination with other cytokines, it acts upon; fibroblasts, myeloid progenitor cells, T-cells, B-cells, and hepatocytes.
    M6518 Macrophage Colony-Stimulating Factor (M-CSF) Human 0.1
    to
    3.0
    ng/mL    		 5  µg
    > 97% pure by (SDS-PAGE) and N-terminal analysis.,
    37 kDa homodimer; each monomer contains 159 amino acids including the N-terminal methionine.
    Endotoxin tested
    Proliferative activity tested using cytokine-dependent monocytic M-NFS-60 cells.

    		 Primarily targets  macrophages and stimulates multiple responses, including; proliferation, cytokine and inflammatory modulator release, cytotoxicity and pinocytosis.
    Also, supports osteoclast differentiation and may act on placental trophoblasts.
    M9170 Macrophage Colony-Stimulating Factor (M-CSF) Mouse
    		 0.3
    to
    3.0
    ng/mL    		 5  µg
    > 97% pure by (SDS-PAGE)
    52 kDa homodimer; each monomer contains 231 amino acids including the N-terminal methionine.
    Endotoxin tested
    Proliferative activity tested using cytokine-dependent monocytic M-NFS-60 cells.

    		 Primarily targets  macrophages and stimulates multiple responses, including; proliferation, cytokine and inflammatory modulator release, cytotoxicity and pinocytosis.
    Also, supports osteoclast differentiation and may act on placental trophoblasts.
    S7901 Stem Cell Factor (SCF) Human;
    recombinant host:
    Escherichia coli    		 2.0
    to
    5.0
    ng/mL    		 10  µg
    > 97% pure by (SDS-PAGE)
    Endotoxin tested
    Proliferative activity tested using TF-1 cells.

    		 Broad activities involving hematopoietic, pigment and primordial germ cell lineages.
    A potent synergistic co-stimulating growth factor which increase the size, but not the differential, of myeloid, erythroid, and lymphoid lineage colonies.
    T4309 Thrombopoietin (TPO) Human;
    recombinant host:
    NSO cells    		 0.4
    to
    4.0
    ng/mL    		 5  µg
    > 97% pure by (SDS-PAGE)
    Predicted 35 kDa from sequence 332 amino acids: glycosylated recombinant product has apparent MW of 82 kDa by (SDS-PAGE).
    Endotoxin tested
    Proliferative activity tested using MO7e cells.

    		 Primary regulatory factor for megakaryocytopoiesis and thrombopoiesis.
    Regulates the growth and maturation of megakaryocytes and their progenitors.
    T4184 Thrombopoietin (TPO) Mouse;
    recombinant host:
    NSO cells    		 0.4
    to
    4.0
    ng/mL    		 5  µg
    >  97% pure by (SDS-PAGE)
    Predicted 35 kDa from sequence 335 amino acids: glycosylated recombinant product has apparent MW of 75 kDa by (SDS-PAGE).
    Endotoxin tested
    Proliferative activity tested using MO7e cells.

    		 Primary regulatory factor for megakaryocytopoiesis and thrombopoiesis.
    Regulates the growth and maturation of megakaryocytes and their progenitors.
    back to top

    help | privacy | technical library | search | home
    terms and conditions | contract manufacturing

    		 
     

    © 2003 Sigma-Aldrich Co. Reproduction forbidden without permission.
    Sigma-Aldrich brand products are sold exclusively through Sigma-Aldrich, Inc. Best viewed in IE5 or higher

    		  

    Order Center Technical Service Customer Service Sigma-Aldrich Search Order Center Technical Service Customer Service Sigma-Aldrich Search